A quick startup guide for using the stopped flow instrument
Important: The user manual by Applied Photophysics is a very extensive and constitutes a detailed
guide for operating and using the Stopped flow instrument. We highly recommend that you read it
before operating the instrument.
An accessible copy of the manual is in PDF version on the computer desktop in PDF form
1. Switch on the lamp power supply and ignite the lamp (red button)
The lamp needs to warm up for 30 minutes
It has a life time of about 2000h but each time one turns it on it decrease its life span
Make sure not to turn off the lamp if other users are expected to use the instrument at the
same day
2. Switch on the Electronic unit box
3. Open nitrogen/compressed air balloon and adjust the pressure to 120PSI (there is a mark on the
pressure regulator)
Note: there are two paths for measuring fluorescent based reactions and one path for measuring
absorbance based reactions. The optical path is 2mm and 10mm for the fluorescent and absorbent
based reactions, respectively. Currently the instrument is configured to measure Absorbance and
fluorescence without the need to change the position of the optical cell house
4. Load each of your samples into 2.5ml syringe (1ml needed from each molecule, concentration in
between the nano to micro molar range).
5. There are 4 positions for loading samples enabling double measurements. Load your syringes on
the two left positions
6. Move the syringe valves to the load position. Press and raise the samples 3 to 4 times to remove
bubbles.
7. Make sure that the syringe pistons are in contact with the single mix drive ram
Syringe
Valve
Valve
load
Syringe
pistons
Single mix
drive ram
Valve
drive
Note! Experiment volume in the flow cell is 20ul
To push it out (remove previous fluids from the flow cell one needs to drive 120ul
It takes about a millisecond to drive the solution into the flow cell o any reaction faster than that can’t
be measured.
8. Start the Software “Pro data SX”
9. For measurements press the “pro data viewer” icon on the software upper ribbon
To view data only one can press the “pro data viewer” icon on the desktop
10. Create a new directory and press “set working directory”
11. Choose signal (Absorbance/ fluorescence)
12. Choose wanted wavelength and press enter
13. Press the “reference” button
14. Load sample: change the valves to the “drive” position
15. Press the drive 3-4 time so as to fill the tubing with your solutions and replace the sample in the
flow cell
16. Trigger “external”
17. Time base (how long to record and how many time points) In the dye mixing demo we used 0.5
and 1000, respectively)
18. Check the repeat box and insert 5 (so it will repeat the measurement 5 times)
19. Acquire
Note: if noises appear or there is a too large variation between the repeated signal one can check the
“Pressure hold” box . Note that shaking the table will cause interference in the data.
For analysis
Choose the sensograms you want
Choose the reaction model and analyze
After work:
Flush the system with water
For long storage flush with 10% ETOH
Switch off the lamp and the electronic unit